Modular platform for multi-tissue integrated cell culture

ABSTRACT

The systems and methods disclosed herein are generally related to a cell culture system. More particularly, the systems and methods enable the culturing and interconnecting of a plurality of tissue types in a biomimetic environment. By culturing organ specific tissue types within a biomimetic environment and interconnecting each of the organ systems in a physiologically meaningful way, experiments can be conducted on in vitro cells that substantially mimic the responses of in vivo cell populations. In some implementations, the system is used to monitor how organ systems respond to agents such as toxins or medications. The system enables the precise and controlled delivery of these agents, which, in some implementations, enables the biomimetic dosing of drugs in humans to be mimicked.

CROSS-REFERENCE TO RELATED PATENT APPLICATIONS

This application claims priority from Provisional U.S. PatentApplication 61/675,688, filed Jul. 25, 2012, which is incorporatedherein by reference in its entirety.

GOVERNMENT LICENSE RIGHTS

This invention was made with government support under W911NF-12-2-0039awarded by the Army Research Office. The government has certain rightsin the invention.

BACKGROUND OF THE DISCLOSURE

In vitro models of human tissue are typically cultured as singlecultures in isolated environments. The isolation of the tissue culturesremoves the interplay between the tissue cultures that is present in invivo systems. The isolated tissue environments make it difficult tostudy systemic issues, such as drug dosing, in in vitro cultures.

SUMMARY OF THE DISCLOSURE

According to one aspect of the disclosure, a modular device forculturing cells includes a control plate, a first and a second cellculture vessel, and an actuator. The first and second cell culturevessels are reversibly coupled to the control plate and are alsorespectively configured to culture a first type of cells and a secondtype of cells. The actuator is within the control plate and configures afluid flow between the first and second cell culture vessels.

In some implementations, the control plate defines a first set ofchannels to route the fluid flow to at least one of the first and secondcell culture vessels. In certain implementations, the cell culturingdevice also includes a fluid routing plate, which is configured toreversibly receive the first and second cell culture vessels. The fluidrouting plate is also configured to couple the first and second cellculture vessels to the control plate. The fluid routing plate defines asecond set of channels to route the fluid flow from the control plate tothe first and/or second cell culture vessel.

In some implementations, the first and second cell culture vessels eachinclude a fluid inlet port and a fluid outlet port to reversibly andfluidically couple to respective channels in the fluid routing plate. Inyet other implementations, the actuator is configured to control a valvewithin the fluid routing plate. The actuator controls at least one ofthe route and flow rate of the fluid flow through the fluid routingplate. In certain implementations, the actuator is configured to closethe valve by deforming a control membrane within the valve.

In some implementations, the actuator is configured to introduce orwithdraw a predetermined amount of an agent, such as a medication or atoxin, into the fluid flow between the first and second cell culturevessels. In some implementations, the first type of cells corresponds toa first organ system and the second type of cells corresponds to asecond organ system. The organ systems are one or a kidney, a lung, anda liver. In some implementations, the first and second organ systems aredifferent.

In some implementations, the actuator is one of a pneumatic actuator, anelectro-mechanical actuator, and a valve. In certain implementations,the control plate includes a plurality of sensors to measure a parameterin the first and second cell culture vessels.

In some implementations, the first and second cell culture vessels aredisposable and the control plate is configured to withstand asterilization process. In yet other implementations, the first andsecond cell culture vessels each include a membrane to support aplurality of cells.

According to another aspect of the disclosure, a method for culturing aplurality of cells includes providing a first cell culture vesselconfigured to culture a first cell type, providing a second cell culturevessel configured to culture a second cell type, and providing a controlplate. The control plate is configured to reversibly couple to the firstand second cell culture vessels and to also control a first fluid flowbetween the first and second cell culture vessels.

In some implementations, the method also includes disposing a firstplurality of cells of the first cell type into the first cell culturevessel and disposing a second plurality of cells of the second cell typeinto the second cell culture vessel. In some implementations, the methodfurther includes coupling the first and second cell culture vessels tothe control plate, and then flowing the fluid between the first cellculture vessel and the second cell culture vessel along the first fluidpath.

In yet other implementations, the method includes reversibly coupling athird cell culture vessel, which is configured to culture a third typeof cell, to the control plate. In some of these implementations, themethod also includes activating an actuator coupled to the control plateto form a second fluid path to the third cell culture vessel.

In some implementations, the first fluid flow includes a liquid flow andthe second fluid flow includes a gas flow. In certain implementations,the first fluid path couples the first cell culture vessel to the secondcell culture vessel and the second fluid path couples the second cellculture vessel to the third cell culture vessel.

In some implementations, prior to coupling the third cell culture vesselto the control plate, the method includes decoupling at least one of thefirst and second cell culture vessels from the control plate andmodifying the first fluid path.

In yet other implementations, the method includes setting a state of anactuator incorporated into the control plate to control at least one ofthe route and flow rate of the fluid flow between the first cell culturevessel and the second cell culture vessel. In some implementations, themethod includes injecting or withdrawing, by an actuator, apredetermined amount of an agent, such as a medication or a toxin, intoat least one of the first cell culture vessel and the second cellculture vessel.

In certain implementations, the first cell type corresponds to a firstorgan system and the second cell type corresponds to second organsystem. In some implementations, the organ systems correspond to one ofa liver, a kidney, and a lung, and, in certain implementations, thefirst and second organ systems are different.

In some implementations, the method includes measuring at least oneparameter within the first and/or second cell culture vessel. In someimplementations, an agent is delivered into at least one of the firstcell culture vessel and the second cell culture vessel and then aresponse to the agent is measured.

According to yet another aspect of the disclosure, a system forculturing cells includes a disposable fluid routing plate. The fluidrouting plate includes a plurality of fluid channels and at last tworeceptacles configured for removably accepting respective first andsecond cell culture vessels. The at least two receptacles are alsoconfigured to couple the first and second cell culture vessels torespective channels of the plurality of fluid channels. The fluidrouting plate also includes at least one valve coupled to at least twoof the fluid channels. The at least one valve is configured to control apath of a fluid flow through the plurality of fluid channels and form afluid path between the at least two receptacles. The system alsoincludes a reusable control plate configured for coupling to the fluidrouting plate. The control plate also includes an actuator forcontrolling the valve in the fluid routing plate.

In some implementations, the system also includes a computer processorcoupled to the control plate and configured to output control signals tothe control plate to configure the position of the actuator. In someimplementations, the system also includes a sensor for measuring aparameter of a cell culture disposed in one of the first and second cellculture vessels. In other implementations, the system includes acomputer processor coupled to the sensor and configured to receive datafrom the sensor.

In yet other implementations, the system includes a fluid pump coupledto one of the control plate and the fluid routing plate to generate afluid flow along the fluid path between the at least two receptacles. Insome implementations, at least one of the receptacles is configured toreceive a plurality of types of modular cell culture vessels configuredfor culturing different respective cell types. In certainimplementations, the system also includes an actuator configured toinject or withdraw a predetermined amount of an agent, such as amedication or a toxin, into the plurality of fluid channels.

BRIEF DESCRIPTION OF THE DRAWINGS

The skilled artisan will understand that the figures, described herein,are for illustration purposes only. It is to be understood that in someinstances various aspects of the described implementations may be shownexaggerated or enlarged to facilitate an understanding of the describedimplementations. In the drawings, like reference characters generallyrefer to like features, functionally similar and/or structurally similarelements throughout the various drawings. The drawings are notnecessarily to scale, emphasis instead being placed upon illustratingthe principles of the teachings. The drawings are not intended to limitthe scope of the present teachings in any way. The system and method maybe better understood from the following illustrative description withreference to the following drawings in which:

FIG. 1 is a schematic of a cell culture system.

FIG. 2 illustrates a schematic of an example cell culture platform thatcan be used in the cell culture system of FIG. 1.

FIGS. 3A and 3B illustrate solid models of an example cell cultureplatform that can be used in the cell culture system of FIG. 1.

FIGS. 4A-4C illustrate solid models of example control plates that canbe used in the cell culture system of FIG. 1.

FIGS. 5A and 5B illustrate solid models of example fluid flow platesthat can be used in the cell culture system of FIG. 1.

FIGS. 6A-6D illustrate example configurations of cell culture vesselsthat can be used in the cell culture system of FIG. 1.

FIGS. 7A and 7B illustrate solid models of an example cell culturevessel.

FIG. 8 illustrates a solid model of an example cell culture vessel.

FIG. 9A illustrates a schematic of an example actuator that can be usedin the cell culture system of FIG. 1.

FIG. 9B illustrates a schematic of an example implementation of anactuator configured to inject and withdraw fluid samples that can beused in the cell culture system of FIG. 1.

FIG. 10 illustrates a flow chart of an example method for culturingcells in the cell culture system of FIG. 1.

FIG. 11 illustrates a schematic of an example use case for the cellculture system of FIG. 1.

DETAILED DESCRIPTION

The various concepts introduced above and discussed in greater detailbelow may be implemented in any of numerous ways, as the describedconcepts are not limited to any particular manner of implementation.Examples of specific implementations and applications are providedprimarily for illustrative purposes.

The systems and methods disclosed are generally related to a cellculture system. More particularly, the systems and methods enableculturing and interconnecting a plurality of tissue types in abiomimetic environment. By culturing organ specific tissue types withina biomimetic environment and interconnecting each of the organ systemsin a physiologically meaningful way, experiments can be conducted on invitro cells that substantially mimic the responses of in vivo cellpopulations. In some implementations, the system is used to monitor howorgan systems respond to agents such as toxins or medications. Thesystem enables the precise and controlled delivery of these agents,which, in some implementations, allows the biomimetic dosing of drugs inhumans to be mimicked.

FIG. 1 illustrates a cell culture system 100. The cell culture system100 includes a cell culture platform 102 within an incubator 104. Thecell culture system 100 also includes a plurality of sensors 106 and amicroscope 108 to monitor the cells within the cell culture platform102. A control computer 110 uses a controller 112 to control the flow offluids and gases through the cell culture platform 102. The fluid flowand gas flow is caused by at least one fluid pump 114 and at least onegas pump 116, respectively. Prior to flowing through the cell cultureplatform 102, fluid is stored in a fluid reservoir 118 and responsive toflowing through the cell culture platform 102 the fluid is stored in awaste reservoir 120.

As described above, the cell culture system 100 includes a cell cultureplatform 102. The cell culture platform 102 and its components aredescribed further in relation to FIGS. 2-9, but briefly, the cellculture platform 102 is a modular platform for culturing cells and/ortissue. As discussed below, the cell culture platform 102 includes acontrol plate, a fluid flow plate and a plurality of cell culturevessels. In some implementations, the control plate is reusable andincludes actuators, valves and sensors used in the culture andmonitoring of cells. In some implementations, the fluid flow plateand/or the cell culture vessels are disposable.

The cell culture platform 102 is housed within an incubator 104. Theincubator 104 maintains an environment within the cell culture platform102 that is conducive for the culturing of the cells and/or tissue. Insome implementations, the incubator 104 controls and/or maintains apredetermined temperature, humidity, carbon dioxide level, oxygen level,or any combination thereof. For example, the incubator 104 may beconfigurable to maintain conditions within the cell culture platform 102that mimic conditions within the human respiratory system. In anotherexample, the incubator 104 is configured to maintain standard cellculture environments, as outlined by a cell culture protocol. Forexample, the incubator 104 can maintain a temperature between about 32°C. and about 37° C. with humidity between about 50% and about 100%. Insome implementations, the incubator 104 removes off gases generated bythe cells within the cell culture platform 102. The incubator 104 alsoincludes a plurality of access ports (not illustrated). The ports allowsensor connections, flow lines, and other lines to pass from the outsideenvironment to the interior of the incubator 104 without affecting thecontrolled environment within the incubator 104.

In some of these implementations, the cell culture system 100 does notinclude a standalone incubator 104. In those implementations, the cellculture vessels of the cell culture platform 102 are reversibly sealedand include heating and other elements that maintain an appropriateenvironmental condition within each cell culture vessel.

The cell culture system 100 also includes a plurality of sensors 106. Insome implementations, one or more of the sensors 106 described hereinare housed within (or a component of) the cell culture platform 102. Afurther description of the sensors 106, including their use andplacement, is described below. In brief, the sensors 106 can be used tomonitor one or more parameters within the cell culture platform 102. Forexample, the sensors 106 can monitor biomarkers, flow rates, pressures,temperatures, gas compositions (e.g., oxygen and carbon dioxide levels),chemical compositions (e.g., drug, toxin and metabolite concentrations),pH levels, electrical parameters (e.g., trans-epithelial electricalresistance) or any combination thereof. In some implementations, thesensors are used for feedback by the control computer 110 in controllingsystem parameters (e.g, environmental conditions) within the cellculture platform 102 and/or incubator 104.

Also as illustrated in FIG. 1, the cell culture system 100 includes amicroscope 108. In some implementations, at least a portion of the cellculture platform 102 is configured to allow visual inspectional of thecells and/or tissue within the cell culture platform 102. For example,the components of the cell culture platform 102 are manufactured fromsubstantially clear materials and/or include view ports. The microscope108 is used to view cells and/or tissue cultured in the cell cultureplatform 102. In some implementations, the microscope 108 is configuredto record still or moving images of the cells and/or tissue within thecell culture platform 102. In some implementations, the microscope 108is an optical light microscope, confocal microscope, fluorescentmicroscope, or, in general, any type of microscope used in the field ofcellular imaging and analysis.

The cell culture system 100 further includes a control computer 110 anda controller 112. In general the control computer 110 controls thecomponents described herein of the cell culture system 100. In someimplementations, the control computer 110 is a general purpose computingdevice. For example, the control computer 110 can be a laptop, tabletcomputer, or smartphone. In other implementations, the control computer110 is a special purposed computer device and includes one or moreprocessors and at least one computer readable medium, such as a harddrive, compact discs, or other storage device. Processor executableinstructions are stored on the computer readable medium. When executed,the instructions cause the control computer 110 to perform the functionsand methods described herein. For example, the control computer 110controls the flow of a fluid into and out of the cell culture platform102 by controlling fluid pumps 114. As described above, in someimplementations the control computer 110 receives data from theplurality of sensors 106 and maintains system conditions responsive tothe received data. The control computer 110 stores the sensor and otherdata on the computer readable medium in response to a request from auser. In some implementations, the control computer 110 enables a userto set specific system parameters through a user interface.

The control computer 110 interfaces with the other components of thecell culture system 100 through a controller 112. In someimplementations, the controller 112 is a component of the controlcomputer 110 or the cell culture platform 102, and is implemented ashardware and/or software. In other implementations, the controller 112is a standalone device that interfaces with the control computer 110 andvarious components of the cell culture system 100 through USB, Firewire,or a similar connection.

The controller includes a plurality of inputs and a plurality of outputsthrough which it interfaces with the various components of the cellculture system 100. The plurality of inputs and outputs of thecontroller 112 can be digital and/or analog inputs and outputs. In someimplementations, the controller 112 includes at least one processor.Using the at least one processor, the controller 112 preprocesses inputsprior to transmitting the input to the control computer 112. Forexample, the controller 112 may “pre-filter” or compress sensor databefore transmitting the sensor data to the control computer 110. In yetother implementations, instructions are loaded onto the controller 112such that the controller 112 can control the cell culture system 100without instruction from the control computer 110. In someimplementations, the controller 112 and/or computer 110 alert a userwhen the cell culture system 100 behavior deviates from predeterminedranges. For example, the control computer 110 may send an alert to theuser when the control computer 110 detects a temperature drop in theincubator 104.

Referring again to FIG. 1, the cell culture system 100 includes at leastone fluid pump 114 and at least one gas pump 116. The fluid pump 114 andthe gas pump 116 (collectively referred to simply as pumps) flow liquidsand/or gases into and through the cell culture platform 102. Extra fluidis stored within the fluid reservoir 118 and can be deposited into awaste reservoir after flowing through the cell culture platform 102. Inother implementations, the fluid is recirculated through the cellculture platform 102. As illustrated, the pumps are independent from thecell culture platform 102. As described below, in some implementations,the pumps are housed within the cell culture platform 102. The pumps caninclude peristaltic pumps, syringe pumps, a series of actuators (i.e.,pneumatic pumps), or any combination thereof. In some implementationsthe pumps are configured to produce a smooth flow, pulsatile flow,periodic flow, or any combination thereof through the cell cultureplatform 102. In yet other implementations, the pumps are directionaland can serve as one way valves within the cell culture platform 102.For example, one way pumps can be included within the cell cultureplatform 102 to force flow in a predetermined manner and not allowbackflow during a pulsatile flow.

The foregoing pumps flow a fluid through the cell culture platform 102and into the below described cell culture vessels. Example fluidsinclude growth medium (or other fluids for cellular growth andsustenance), test agents, toxins, medicaments (e.g., antibiotics,vaccines, biologics, and medical countermeasures), or any combinationthereof. In some implementations, the pumps are configured to induce apredetermined shear force on the cells within the cell culture platform102. The shear force may be selected to mimic physiological conditionsor for experimental purposes. For example, epithelial cells may formmore physiologically representative cellular barriers when culturedunder an appropriate shear force. In some implementations, the flowrates at which the pumps flow fluid are selected to mimic blood flowrates typically seen in parts of the circularity system.

Referring now to FIG. 2, FIG. 2 is a schematic illustrating componentsof the cell culture platform 102. The individual components of the cellculture platform 102 are described in detail in relation to FIGS. 4-9.As a brief introduction, the cell culture platform 102 includes acontrol plate 202, a fluid flow plate 204, and a plurality of cellculture vessels 206(1)-(n). The fluid flow plate 204 is coupled to thecontrol plate 202, and a plurality of cell culture vessels 206(1)-206(n)are coupled atop the fluid flow plate 204. The cell culture platform 102further includes a plurality of fluid and/or gas inlet/outlet ports 208.As illustrated, the ports 208 are components of the control plate 202.In other implementations, the control plate 202, fluid flow plate 204,and/or cell culture vessels 206(1)-206(n) each include one or more ports208.

Continuing the cell culture platform 102 overview, the cell cultureplatform 102 is used to culture cells and/or tissues. In someimplementations, this includes the culture of multiple types of cellsand/or tissue from different organ systems. In some implementations, asdescribed below, the cell culture vessels 206 are configured to include3-dimensional cell culture scaffolds to support and culture the cellsand/or tissues. The remaining plates of the cell culture platform 102facilitate interaction (e.g, fluidic communication) between thecells/tissues cultured within the cell culture vessels 206(1)-206(n),and enable the cell culture vessels 206 to be interconnected inphysiologically meaningful ways.

In some implementations, the components of the cell culture platform 102are reversibly coupled to one another. For example, the components ofthe cell culture platform 102 can be coupled to one another with claps,screws, via vacuum, adhesive or any combination thereof. In someimplementations, the coupling element (e.g, a screw) that is used tocouple the cell culture vessel 206 to the fluid flow plate 204 passesthrough the fluid flow plate 204 to also couple the fluid flow plate 204to the control plate 202.

In certain implementations, one or more of the components of the cellculture platform 102 are disposable and/or reusable. For example, thecontrol plate 202 may house control connections to the controller 112,sensor connections, actuators, custom components, or any combinationthereof is intended to be reused with disposable fluid flow plates 204and disposable cell culture vessels 206.

In some implementations, the disposable elements include passivestructures that are produced using low-cost processes such as machining,injection modling, or embossing. In some implementations, these passivestructures are controlled via actuators within the control plate 202. Insome implementations, the control plate 202 provides a foundation towhich disposable fluid flow plates 204 and cell culture vessels 206 maybe modularly added.

FIG. 3A is solid model illustrating cell culture platform 102 in greaterdetail. As illustrated, eight cell culture vessels 206 are coupled to afluid flow plate 204, which is, in turn, coupled to a control plate 202.The control plate 202 includes a first type of cell culture vessel206(a) and a second type of cell culture vessel 206(b).

FIG. 3B is a side view of the model from FIG. 3A illustrating the cellculture platform 102. In some implementations, the cell culture vessels206 are sealed with reversibly coupled lids 302 and 306. The lid 302includes a port 304, which in some implementations, is used to flowgases and/or liquids into the cell culture vessel 206(b). The lid 306 isa sealed lid and does not include a port. As illustrated, the cellculture vessels 206 are coupled to the fluid flow plate 204 with screws308.

Below, each of the control plate 202, the fluid flow plate 204, and thecell culture vessels 206 of FIGS. 2, 3A, and 3B are described in turnand in greater detail with reference to FIGS. 4-9.

As set forth above in reference to FIGS. 2, 3A, and 3B, the cell cultureplatform 102 includes a control plate 202. In general, the control plate202 contains reusable connectors, actuators, and/or sensors thatinterface with the fluid flow plate 204 and/or cell culture vessels 206.In some implementations, the placement of the connectors, actuatorsand/or sensors in the reusable control plate 202, provides a costsavings as portions of the cell culture platform 102 that directlyinteract with cells can be disposed of after experimentation, while themore expensive components can be reused. As described below, in someimplementations, the control plate 202 is manufactured from a plastic ora multi-layer printed circuit board.

In some implementations, the control plate 202 includes between 5 and10, between 10 and 30, between 30 and 50, between 50 and 100, or between100 and 200 actuators. The actuators are used to control fluid flowthrough the fluid flow plate 204 and/or cell culture vessel 206, and, insome implementations, are used as pumps. The actuators control fluidflow by activating valves within the control plate 202, fluid flow plate204 and/or cell culture vessel 206. In implementations where theactuators are configured as pumps, they pump between about 100 nL andabout 500 nL, between about 500 nL and 1000 nL, or between about 1000 nLand about 2000 nL/min of fluid through a channel. The flow induced bythe actuator pumps can have a continuous, single shot, and/orreciprocating flow profile.

In some implementations, the pump is configured to inject apredetermined dosage of a toxin, test agent, medicaments (e.g.,antibiotics, vaccines, biologics, and medical countermeasures), or anycombination thereof into the fluid flow plate 204 and/or the cellculture vessel 206. For example, on a predetermined cycle (e.g., onceper day, three times a day, once per hour, etc.) the pump-configuredactuator may be configured to deliver an insulin dose to a cell culturevessel containing liver cells. In some implementations, apump-configured actuator withdraws a predetermined fluid sample volumefrom the fluid flow plate 204 and/or the cell culture vessel 206. Forexample, the actuator may withdraw 100 nL from a cell culture vesselevery hour, such that a medicament, analyte, or toxin, or otherbiologically relevant material concentration can be determined in thecell culture vessel.

In various implementations, the actuators are pneumatic actuators,electromagnetic actuators, valves, or a combination thereof. Themechanism of the actuator activation is described further in relation toFIG. 9A, and the mechanism of the actuator when acting as a pump toinject or withdraw fluid samples is described in relation to FIG. 9B.Briefly, the actuators include a membrane, which is driven by a piston.When activated, the actuator drives the piston and membrane into achannel placed above the actuator. The membrane shunts the flow of afluid through the channel. In some implementations, pneumatic actuatorsare used because in some implementations, the activation of anelectromagnetic actuator may induce heat or electromagnetic noise thatmay interfere with certain sensor applications such as transepithelialelectrical resistance.

The actuators enable customized control of fluids through the cellculture platform 102. The use of a membrane in the actuator enablesseparation of biological liquids from the reusable components of thecontrol plate 202. In some implementations, the flexible membrane usedin the actuator (and/or pump structures) is manufactured from, but isnot limited to, polyimide- and polyurethane-based materials. In someimplementations, substantially the entire, or at least large portionsof, the top surface of the control plate 202 is covered with themembrane.

In some implementations, the control plate 202 includes a fixed formfactor that couples (or mates) with the cell culture vessel 206 and/orthe fluid flow plate 204. As described below, fluid flow plate 204 andcell culture vessel 206 can be configured differently responsive to theneeds an experiment. In these implementations, the standardized formfactor of the control plate 202 enables the mixing and matching of othermodular components to the control plate 202.

As introduced above, the control plate 202 includes one or more sensors106 and/or sensor connections. For example, the control plate 202 caninclude flow meters, gas sensors, pH sensors, temperature sensors,transepithelial electrical resistance (TEER) sensors, or any combinationthereof. In some implementations, the flow sensor is a thermal flowsensor. In certain implementations, the sensors 206 are mounted topolyimide substrates and separated from fluids by the above describedmembrane.

In implementations including sensor connections (or sensor expansionports) the sensors 106 described herein are added to the control plate202 based on the requirements of an experiment. For example, aresearcher conducting a flow experiment may choose to only attach flowsensors to the control plate 202 and may forgo other sensors such as aph sensor. In some implementations, removing sensors 106 by decouplingthem the from the expansion ports, facilitates the reusability of thecontrol plate 202 by enabling delicate components of the control plate202 to be removed prior to sterilization of the control plate 202. Insome implementations, the sensor expansion ports are input/output portsfor the controller 112, and allow for the connection of custom sensorsto the control plate 202.

In some implementations, the control plate 202 includes at least oneheating element. The heating element is employed to maintain aconfigurable temperature within one or more of the cell culture vessels206. In some implementations, use of a heating element and closed cellculture vessels 206 enable experiments to be conducted without anincubator 104, as a predetermined microcondition can be maintainedwithin each cell culture vessel 206.

In yet other implementations, the control plate 202 includes anauxiliary agent delivery module. The module connects to the controlplate and enables specific agent dosage to one or more of the cellculture vessels 206.

To further describe the control plate 202 discussed above, FIGS. 4A-4Cillustrate example implementations of the control plate 202. A person ofordinary skill in the art will recognize that features of the variouscontrol plates described below may be applied to any of the othercontrol plates described herein.

FIG. 4A is a top view illustrating a pneumatic control plate 400. Thecontrol plate 400 includes a plurality of actuators 402 to act on flowchannels within the fluid flow plate 204. The control plate 400 alsoincludes a plurality of pneumatic ports 404 to control the plurality ofactuators 402. A fluid flow enters the control plate 400 at inlet port408 and exits the control plate 400 through the plurality of flow ports406(a) and 406(b). The control plate 400 further includes a plurality ofview ports 410 that provide optical access to the underside of the cellculture vessels 206.

As described above, the control plate 400 includes a plurality ofpneumatic actuators 402. As illustrated, control plate 400 includestwenty actuators divided into four actuator groups 412. Each actuatorgroup 412 corresponds to the intersection of two channels in the fluidflow plate 204. The actuator 402(a) lies at the center of the actuatorgroup 412, and, when activated, stops the flow through all four branchesof the intersection. Each actuator 402(b)-405(e) controls the flow ofthe fluid into its respective branch of the intersection.

The control plate 400 also includes a plurality of flow ports 406. Asillustrated, the control plate 400 includes a first type of flow port,flow port 406(a), and a second type of flow port, flow port 406(b). Flowport 406(b) has a larger relative diameter compared to flow port 406(a).In some implementations, a larger diameter port 406 enables a greaterrelative volume of fluid to flow through the flow port 406. In someimplementations, each flow port 406 is coupled to a single inlet port408. In other implementations, the control plate 400 is configurable toprovide separate sources to one or more of the flow ports 406. In someimplementations, the opening of the flow port 406 is counter sunk intothe control plate 400 and includes a washer or O-ring in the countersunk area. The washer or O-ring prevents fluid leakage when a fluid flowpasses from the control plate 400 to the fluid flow plate 204.

The control plate 400 also includes four viewing ports 410. The viewingports 410 are pass throughs (or vias) that enable optical access to thedorsal side of the cell culture vessels 206 eventually coupled to thecell culture platform 102. In some implementations, the control plate400, fluid flow plate 204, and/or cell culture vessels 206 aremanufactured from optically clear materials such that cell cultures areoptically accessible without view ports 410. In some implementations,the components of the cell culture system 100 are substantiallyoptically clear and include a plurality of view ports 410.

FIG. 4B is a cross sectional view illustrating the internal flowchannels of the control plate 400. As illustrated, the control plate 400includes the channels 414(1)-414(6). The channel 414(1) corresponds thefluid inlet port 408. The channels 414(2)-414(6) each correspond to oneof the pneumatic ports 404 and act as control channels for the abovedescribed actuators 402(a)-402(e). FIG. 4B illustrates that eachactuator group 412 is connected to the same control channels414(2)-414(6), and thus operate in unison. In some implementations, eachactuator 402 within actuator group 412 the control plate 202 isindividually controllable.

The channel 412(1) includes a plurality of stems to route a fluid to theflow ports 406. The flow port 406(b) includes a relatively largerdiameter compared to the flow port 406(a). Accordingly, stem 416, whichcorresponds to the larger flow port 406(b), includes a larger diameterto support the increased flow through flow port 406(b). In comparison,stem 418, which corresponds to flow port 406(a) includes a relativelysmaller diameter. In some implementations, the stems 416 and 418 and thefluid flow channels described herein have a diameter of about 1-5 mm,about 5-10 mm, and about 15-25 mm.

As described above, in some implementations, the actuator is anelectromagnetic actuator. FIG. 4C is an isometric view of a controlplate 450 with electromagnetic actuators 452. The control plate 450 ismanufactured on a printed circuit board 454, and similar to controlplate 400, includes a plurality of view ports 410. Additionally, thecontrol plate 450 includes a membrane 456 that protects the electronicsof the control plate 450 from the fluids contained in the above layers.The control plate 450 also includes a plurality of electrical connectors458. As illustrated, control plate 450 does not include fluid flowchannels. In this implementation, the fluid inlet port 408 would beincluded within the fluid flow plate 204 and/or cell culture vessels206. In other implementations, the control plate 450 is configured toinclude a fluid inlet port 408 similar to control plate 400.

In some implementations, the electromagnetic actuators enable a smallerrelative footprint compared to the control plate 400. In someimplementations, the actuators 452 are implemented for bi-stableoperation with fixed mechanical stops for the pistons they incorporate.This enables the actuators to have reproducible stroke volumes and onlyrequire power during engaged-unengaged transitions. As suggested above,in some implementations, the control plate 400 with pneumatic actuatorsis used when it is desired to have no, or a reduced number of,electrical components within the cell culture platform 102. For example,if an experimenter is performing electro-physiological experiments andthe electrical components of the control plate 202 interfere with theelectrophysiology recordings, then the experimenter may choose to use apneumatic based system.

The control plate 450 also includes a plurality of connectors 458. Insome implementations, the connectors 458 are used to electrically couplethe control plate 450 to the controller 112 for the purpose ofactivating the actuators 452. In other implementations, the connectors458 are used to connect sensors 106 to the control plate 450 andultimately to the control computer 110. In some implementations,pneumatic implementations also include connectors 458 for the connectionof sensors 106.

Referring back to FIGS. 2, 3A, and 3B, the cell culture platform 102includes a fluid flow plate 204. The fluid flow plate 204 includes aplurality of flow channels defined there through. The fluid flow plate204 acts as an interface between the control plate 202 and the cellculture vessels 206. For example, the fluid flow plate 204 interfaces onits dorsal side with the flow ports 406 of the control plate 400. Afluid flow is then routed from the control plate 202 to the fluid flowplate 204 where the fluid can be routed to the cell culture vessels 206.

In some implementations, the fluid flow plate 204 is constructed fromtransparent, chemically stable, and mechanically robust thermoplasticmaterials such as polystyrene. The material of the fluid flow plate 204is selected to avoid chemical instabilities and chemical absorption.

In some implementations, dynamic control over flow through the fluidflow plate 204 is achieved using the above described actuators of thecontrol plate 202. For example, the user can activate specific actuatorsto close, control the flow rate of, or route fluid away from channels.

In some implementations, the fluid flow plate 204 is disposable. Inother implementations, the fluid flow plate 204 also includes actuators,sensors, and/or “reusable” components as described herein.

Now referring to FIG. 5A, which illustrates an isometric view of anexample fluid flow plate 500. The top surface of the fluid flow plate500 includes a plurality recesses (or mortises) 504. As described below,the cell culture vessels 206 include matching projections (or tenons).The mortises 504 and tenons interlock and properly align cell culturevessels 206 with the flow ports 502. As illustrated, the flow ports 502are included in a subset of the mortises 504. In some implementations,each mortise 504 includes a flow port 502.

As illustrated, and referring back to FIGS. 3A and 3B, the fluid flowplate 500 supports six cell culture vessels 206. In someimplementations, the fluid flow plate 500 supports between 1 and 10, 10and 20, 20 and 50, 50 and 100 cell culture vessels 206.

FIG. 5B illustrates a cross-sectional view of the fluid flow plate 500from FIG. 5A. As revealed by the cross-sectional view, the fluid flowplate 500 includes a plurality of fluid flow channels 508. In someimplementations, the fluid flow channels 508 connect one or more flowports 502 to other fluid flow channels 508, flow ports 406 on thecontrol plate 202, or a combination thereof. Thus, in someimplementations, the fluid flow channels 508 connect one or more cellculture vessels 206, interconnect different portions of a single cellculture vessel 206, and/or connect the fluid flow channels 508 to thecontrol plate 202. In some implementations, the fluid flow plate 500includes a plurality of layers each of which include additional fluidflow channels 508. For example, the fluid flow plate 500 may include afirst layer of fluid flow channels 508 that run along a first axis and asecond set of fluid flow channels 508 that run orthogonal to the firstaxis.

Referring back to FIGS. 2, 3A, and 3B, the cell culture platform 102includes a plurality of cell culture vessels 206(1)-(n), where n is thenumber of cell culture vessels. As described above, various cell cultureplatforms 102 can support between 1 and 10, between 10 and 20, between20 and 50, or between 50 and 100 cell culture vessels 206. In someimplementations, the cell culture vessels 206 are configured to house aspecific cell type and/or cells from a particular organ type. In someimplementations, the cells from the particular organ type include aplurality of cells types related to the particular organ. For example,when the cell culture vessel 206 is configured to house organ cells, thecell culture vessel can be configured to culture Loop of Henle thinsegment cells, tubule cells, collecting duct cells, and glomerulusparietal cells. In some implementations with multiple cells typesrelating to a particular organ type, a first cell type related to theorgan is cultured above a permeable membrane and a second cell typerelated to the organ is cultured below the permeable membrane.

In some implementations, the cell culture vessels 206 include a commonexterior form factor regardless of the internal configuration of thecell culture vessel 206. For example, each cell culture vessel 206 caninclude the above described tenons and fluid ports at predeterminedlocations so the cell culture vessels 206 can be placed in any cellculture vessel slot on the fluid flow plate 202.

In some implementations, the cell culture vessels 206 are configured tosupport specific cell and/or organ tissue types. In someimplementations, the cell culture vessels 206 may include specificscaffolds or structures to enable 3-dimensional cell growth of aspecific cell and/or organ type. In other implementations, the cellculture vessels 206 are configured to support specific cell and/or organtissue types by providing a predetermined flow rate to the cell culturevessel 206 and/or by providing predetermined fluids (e.g., specificmedia mixtures) to the cell culture vessel 206. For example, a cell typethat requires a high shear force can be cultured in a cell culturevessel 206 with a plurality of input ports and a plurality of outputports. The plurality of input and output ports enable a relativelylarger volume of fluid to flow through the cell culture vessel 206, thusimparting a relatively larger shear force on the cells within the cellculture vessel 206. In some implementations, cells that require littleor no shear force may be cultured in cell culture vessels with a singleport, such that nutrients diffuse into the cell culture vessel throughthe single port under no force from a fluid flow.

In other implementations, based on their physiological requirements,cells are cultured in a scaffold submerged in media or on a membrane atan air-liquid interface. For example, alveolar cells from the lung maybe placed in a cell culture vessel 206 that is designed to provide airto the top-side of the cells while supplying the dorsal side of thecells with nutrients. In another example, liver cells may be cultured ona permeable membrane above a reservoir such that diffusion can occurthrough the liver cell layer and membrane to the reservoir.

As described in greater detail below, in some implementations, the cellculture vessels 206 include slots for one or more cell culture inserts.The cell culture inserts house the cells cultured in the cell culturevessel 206. The cell culture inserts are removable and enable theindividual cultures to be seeded and grown outside of the cell culturesystem 100. For example, a company may sell pre-seeded cell cultureinserts, which a researcher purchases and then inserts into a cellculture system 100.

In some implementations, the cell culture vessels 206 include multiplecompartments that are separated by semi-permeable membranes. In someimplementations, the membranes can include specific matrix componentsrepresenting the surface chemistry, mechanical stiffness, and porosityof in vivo tissues. In some implementations, cells are cultured directlyon the membranes.

As with the other components of the cell culture platform 102, in someimplementations, the cell culture vessels 206 are disposable. The cellculture vessels 206 are manufactured from optically transparentmaterials such as polystyrene and/or polyimide. The cell culture vessels206 materials are stable and compatible with cell culture and biologicalfluids relative to conventional microfluidic materials. For example, insome implementations, the cell culture vessels 206 are manufactured fromPDMS. In some implementations, disposable cell culture vessel componentsare manufactured from thermoplastics such as polystyrene, polycarbonate,cyclic olefin copolymer (COC), or any combination thereof. In someimplementations, the cell culture vessels 206 are manufactured by directmachining, embossing, injection molding, or any combination thereof maybe used. In some implementations, the control plate 202 and/or fluidflow plate 204 are manufactured through similar processes with similarmaterials to those described above.

In some implementations, the cell culture vessels 206 and/or the fluidflow plate 204 include one-way valves. The one-way valves enable thecell culture vessels 206 to be temporally removed from the fluid flowplate 204 during experimentation. For example, a user may remove a cellculture vessel 206 from the cell culture platform 102 to perform aseparate experiment or test on the cells within the removed cell culturevessel 206.

In some implementations, the above described fluid reservoir 118 and/orwaste reservoir 120 can have the same form factor as a cell culturevessel 206, enabling the fluid reservoir 118 and/or the waste reservoir120 to be modularly added to the cell culture platform 102. The fluidflow plate 202 and the control plate 202 can then flow growth media orother fluids (such as a medication or toxin) from the reservoir to theother components of the cell culture platform 102.

As described below, in some implementations, the cell culture vessels206 include customized scaffold structures for each physiological systemmodel. In some implementations, the scaffolds (also referred to as cellculture inserts) enable individual models to be developed separatelyfrom the cell culture platform 102 and then supplied individually forpractical implementation.

In some implementations, specialized drug storage and delivery may berequired for specific cell culture vessels 206 (e.g, delivering insulinto a cell culture vessel 206 culturing liver cells). Theseimplementations can include custom modules fitted to the above describedlids of specific culture wells. For example, and referring to FIG. 3B,the port 304 on lid 302 may be used to enable delivery of an agent tothe interior of cell culture vessel 206(b). In some implementations, thedelivery module is controlled by the control plate 202 and/or directlyby the controller 112.

FIGS. 6A-6D illustrate schematics of various example cell culturevessels. As illustrated, each cell culture vessels 600, 610, 620, and630 includes an inlet port 602 and an outlet port 604. In someimplementations, the cell culture vessels include a plurality of inletports 602 and/or a plurality of outlet ports 604. In certainimplementations, each port of a cell culture vessel 206 is configured tobe an inlet pot 602 or an outlet port 604 by configuring the fluid flowplate 204 with the one or more actuators in the control plate 202.

Each cell culture vessel 600, 610, 620, and 630 also includes a cellculture insert 606. As described above, the cell culture insert 606enables the off-platform culturing of cells. The cell culture vesselsinclude slots which secure the cell culture inserts 606 in place. Insome implementations, the bottom surface of the cell culture insertincludes a semi-permeable membrane on which cells are cultured.

FIG. 6A illustrates a cell culture vessel 600 configured for a basalflow 608. As described above, some cells are responsive to specificflows and/or shear forces. For example, a cell population of liver cellsmay more closely mimic in vivo liver cells if exposed to a shear force.By employing a cell culture insert 606 with a permeable membrane, theconfiguration of cell culture vessel 600 exposes a cell's basal membraneto a flow and thus the described shear force. In some implementations, abasal flow allows the dorsal surface to be exposed to gases. Forexample, this type of configuration may be used to mimic alveolartissue. In this example, alveolar epithelial cells are cultured in thecell culture insert 606. Nutrients are supplied to the cells through thebasal flow 608, as the cells are exposed to gas along their top surface.

FIGS. 6B and 6C illustrate cell culture vessels 610 and 620,respectively. The cell culture vessels 610 and 620 are configured toprovide a top flow. The cell culture vessel 610 includes a raised cellculture insert 606. The raised cell culture insert 606 enables diffusionthrough the cells and into a reservoir space 611 located beneath theinsert 606(b). In some implementations, the cell culture configurationof cell culture vessel 620 is used to culture gut epithelial cells. FIG.6D illustrates the cell culture vessel 630. The cell culture vessel 630is configured to allow flow above and below the cell culture insert 606.

FIG. 7A illustrates an isometric view of one example implementation of acell culture vessel 630, similar to the cell culture vessel 206(b) inFIG. 3A. Exteriorly, each wall of the cell culture vessel 700 includes arecess used to secure the cell culture vessel 700 to a fluid flow plate204 with thumb-screws. The interior of the cell culture vessel 700includes a top flow area 704 and cell culture area 706. In someimplementations, the floor of the cell culture area 704 is asemi-permeable membrane.

FIG. 7B illustrates an isometric cutaway view of the cell culture vessel700. As revealed by the cut-a-way, the cell culture vessel 700 includesa lower flow area 708. Fluid flows into and out of the lower flow area708 through ports 710. The arrow 712 illustrates one possible flowpattern through the cell culture vessel 700. A lid 714 is optionallycoupled to the cell culture vessel 700. The lid 714 is manufactured withsimilar materials as the cell culture vessel 700. In someimplementations, the lid 714 is transparent to provide optical access tothe cells within the cell culture area 706. The lid 714 also includes aplurality of access ports 716. In some implementations, the access ports716 are used to introduce a gas and/or a liquid into the top flow area704. The gas and/or liquid is supplied to the access ports 716 throughthe control plate 202 and/or the fluid flow plate 204 in someimplementations. In other implementations, the gas and/or liquid supplyto the access ports 716 is independent of the cell culture platform 102.In some implementations, the cell culture vessel 700 is used to culturelung tissue. For example, lung cells are cultured within the cellculture area 706. Nutrients in the lower flow area diffuse to the cellsthrough the semi-permeable membrane of the cell culture area 706. Gas,emulating gas within a human's lungs, is passed into the top flow area704 through the access ports 716.

FIG. 8 illustrates another implementation of a cell culture vessel 206.FIG. 8 illustrates a top view of cell culture vessel 800, similar to thecell culture vessel 206(a) in FIG. 3A. The cell culture vessel 800includes an inlet port 802. The fluid flow entering the cell culturevessel 800 is directed around a wall 804 and toward an outlet 806. Theoutlet 806 is recessed within a slot 808, which is similar to abovedescribed slots for securing the cell culture inserts. In the cellculture vessel 800, a portion of the fluid flow flows through the cellsand membrane of the cell culture insert to reach the outlet 806.Recesses 810 enable excess fluid to bypass the cell culture insert andflow directly to the outlet 806. In some implementations, a cell culturevessel similar to the cell culture vessel 800 is used for culturingcells, such as liver cells, in the presence of a shear force.

FIG. 9A illustrates a cross sectional view of an actuator 900 suitablefor inclusion in the control plate for controlling fluid paths in thefluid flow plate. The actuator 900 is housed within control plate 902. Afluid flow plate 904, which includes the flow channel 906, is coupled tothe control plate 902. To close the flow channel 906, the actuator 900drives its piston upward. As described above, a membrane 908 separatesthe actuator from the fluid of the fluid flow plate 904. Once deployedthe piston drives into a recess 910 in the top of the flow channel. Thiscreates a seal, closing the channel 906.

FIG. 9A also illustrates a fluidic capacitor 912. In someimplementations, one or more fluidic capacitors 912 are included in theflow channels of the cell culture platform 102. The fluidic capacitor912 smooths a fluid flow through the channel to which it is attached.The fluidic capacitor 912 includes a membrane 914 above a cavity 916.Responsive to a pulsatile wave (or other non-smooth flow) the membrane914 deforms into the cavity 916. The expansion of the channel into thecavity 916 slows the pulsatile wave and smooths the flow through thechannel.

FIG. 9B illustrates a cross sectional view of example actuatorsconfigured to inject and/or withdraw fluid samples for a cell culturesystem. As illustrated in FIG. 9B, a fluid channel 950 runs below a cellculture vessel 952. An injection/withdrawal (I/W) module 954 is coupledto one end of the channel 950. The I/W module 954 includes a firstactuator 956, which when activated seals the I/W module 954 off from thefluid channel 950. The mechanism of the first actuator 956 is similar tothe above described actuator 908 illustrated in FIG. 9A. Briefly, thefirst actuator 956 drives a membrane 962 into a recess in the top of thefluid channel 950, which creates a seal and closes the I/W module 954off from the fluid channel 950. The I/W module 954 also includes asecond actuator 958, which is coupled to a second membrane 964. The I/Wmodule 954 also includes a reservoir 960 to store a fluids for injectionand/or after withdrawal. In some implementations, the I/W module 954also includes an access port (not illustrated) to enable the injectionand/or withdrawal of fluid from the reservoir 960.

To withdraw (also referred to as sipping) a sample from the fluidchannel 950, the first actuator 954 lowers. With the first actuator 954lowered, a fluid can enter the I/W module 954. The second actuator 958retracts its piston, and drives the second membrane 964 upward. Theupward movement of the membrane 964 creates a vacuum in the reservoir960, which draws a fluid from the fluid channel 950 into the reservoir960. To inject a fluid into the fluid channel 950, a similar processoccurs. During a fluid injection, the second actuator 958 extends itspiston, creating a pressure build up in the reservoir 960. Responsive tothe first actuator 956 opening access to the fluid channel 950, thepressure build up drives the fluid in the reservoir 960 out of the I/Wmodule 954 and into the fluid channel 950.

In some implementations, the I/W module 954 does not require the secondactuator 958 to withdraw fluid from the fluid channel 950. For example,the flow present in the fluid channel 950 may drive fluid into thereservoir 960. In some implementations, the I/W module 954 is acomponent of the above described fluid flow plate, cell culture vessels,or control plate. For example, the I/W module 954 may be a component ofa cell culture vessel and inject or withdraw fluid directly from thecell culture vessel. In other implementations, the I/W module 954 is aseparate module form the cell culture platform, and may be modularlyadded to any of the cell culture vessels and/or the fluid flow plate.

FIG. 10 illustrates a flow chart of a method 1000 for culturing aplurality of cells. In some implementations, the method 1000 is used totest the interplay of organ systems in vitro. The method 1000 includesproviding a first and second cell culture vessel (step 1001). The method1000 also includes providing a cell culture platform (step 1002). Cellsof a first type are disposed in the first cell culture vessel and cellsof a second type are disposed in the second cell culture vessel (step1003). Then, the cell culture vessels are coupled to the cell cultureplatform (step 1004) and a fluid path (also referred to as a fluidcircuit) is configured to the first and/or second cell culture vessels(step 1005). The method 1000 also includes flowing a fluid through thecell culture platform to the first and second cell culture vessels (step1006).

As set forth above, the method 1000 begins with the provision of a firstand second cell culture vessel (step 1001) and cell culture platform(step 1002). The first and second cell culture vessels can be similar tothe cell culture vessels described above in relation to FIGS. 2-3B, and6A-8. In some implementations, the first and second cell culture vesselsare configured differently. For example, the first cell culture vesselcan be configured to culture tissue from a first organ (e.g., lungtissue), and the second cell culture vessel can be configured to culturetissue from a second organ (e.g., liver tissue). For example, the firstcell culture vessel may be the cell culture vessel 700 illustrated inFIG. 7A and the second cell culture vessel may be the cell culturevessel 800 illustrated in FIG. 8. In some implementations, the cellculture platform is the cell culture platform 102 discussed above. Insome implementations, one or more cell culture vessels are alreadycoupled to the cell culture platform 102 prior to the beginning of themethod 1000.

Next, a first type of cells are disposed in the first cell culturevessel and a second type of cells are disposed in the second cellculture vessel (step 1003). In some implementations, the cell culturevessel configurations selected in step 1001 is responsive to the type ofcells a user intends to use in step 1003. In some implementations, auser is able to mimic an organ system by combining a specific cell typewith a specific cell culture vessel configuration. For example, a usermay select to combine alveolar cells with a cell culture vesselconfiguration that provides a liquid-gas interface (e.g, the cellculture vessel 700 illustrated in FIGS. 7A and 7B).

In some implementations, the first and second cell types are differentcell types. In these implementations, a user may combine different celltypes and cell culture vessel configurations to mimic a plurality oforgan systems. In some implementations, the organ systems correspond totwo or more of a liver, a lung, or a kidney. As described below, in someimplementations the modular combination of multiple organ systemsenables a user to study the interactions between those organ systems. Inother implementations, a user can use a cell culture platform culturinga plurality of interconnected organ systems to study drug dosing anddrug uptake.

Next, the first and second cell culture vessels are coupled to the cellculture platform (step 1004). In some implementations, as describedabove in relation to FIGS. 2-3B, the cell culture vessels are coupled toa fluid flow plate, which acts as an interface between a control plateand the cell culture vessels. In some implementations, the cell culturevessels are reversibly coupled to the control plate and/or fluid flowplate.

The method 1000 further includes configuring a fluid circuit between thefirst and second cell culture vessels (step 1005). As described above,in some implementations, an actuator is coupled to (or within thecontrol plate). Activation of the actuator controls at least one valvein the fluid flow plate and/or cell culture vessels. By activating theone or more actuators in the cell culture platform, a user configures afluidic circuit that routes the fluid flow between the first and secondcell culture vessels.

Responsive to coupling the first and second cell culture vessels to thecontrol plate, a fluid is flowed through the cell culture platform tothe first and second cell culture vessels (step 1006). In someimplementations, the fluid enters the cell culture platform at aninterface with the control plate and/or the fluid flow plate. In yetother implementations, the fluid enters the cell culture platformthrough one or more of the cell culture vessels. In someimplementations, flowing the fluid through the cell culture platformconstitutes recirculating the fluid through the cell culture platform.In some implementations, the fluid is a growth medium, blood, a gas, orany combination thereof.

In some implementations, the method 1000 further includes disposing athird cell type into a third cell culture vessel and then coupling thethird cell culture vessel to the cell culture platform in addition to orin place of the first and second cell culture vessels. In otherimplementations, the method 1000 also includes reconfiguring the fluidcircuit created in step 1006 by activating one or more actuators. Forexample, by activating one or more of the actuators, the above describedfluid circuit can be reconfigured to include the third cell culturevessel. In other implementations, the method 1000 includes rearrangingand/or removing the first, second, and/or third cell culture vesselswithin the cell culture platform. In yet other implementations, themethod 1000 includes measuring a parameter within the cell cultureplatform 102. For example, a temperature in one of the cell culturevessels and/or a flow rate through the fluid circuit may be measured. Insome implementations, a cell culture vessel is temporally removed fromthe cell culture platform 102 to perform the measurement. In otherimplementations, a cell culture vessel is permanently removed andreplaced with a cell culture vessel housing similar or different cellsor organ tissue type.

One of ordinary skill in the art will recognize that in someimplementations the above method steps of the method 1000 may beperformed in a different order or one or more of the method steps may beomitted. For example in one implementation, the fluid circuit may beconfigured prior to the coupling of the cell culture vessels to the cellculture platform. In a similar example, a user may purchase a fluid flowplate that includes preconfigured fluid flow channels and therefore doesnot have to be configured once coupled to the cell culture platform.

FIG. 11 illustrates an example schematic of a use case of the abovedescribed system. The schematic illustrates a system 1100 that, in someimplementations, is used to investigate drug candidates. The system 1100corresponds to a cell culture platform culturing cells that correspondto four organ systems. In some implementations, one or more cell culturevessels correspond to each organ system. The four organ systems of thesystem 1100 include tracheobronchial tissue 1102, alveolar tissue 1104,small intestine tissue 1106, and liver tissue 1108. Using the pluralityof valves 1110 and valve groups 1112, which correspond to actuators in acontrol plate, two circulatory circuits are created within the fluidflow plate used to implement the system 1100. The first circuit 1114represents a circulatory (or cardiovascular) system. The first circuit1114 provides nutrients to each of the organ systems 1102, 1104, 1106,and 1108. In some implementations, the fluid used in the transport ofnutrients and other chemicals to each of the organ systems 1102, 1104,1106, and 1108 is a growth medium, blood, or a blood analyte. The secondcircuit 1116 (illustrated as a dashed line) is coupled to only the smallintestine tissue 1106 and the liver tissue 1108. The second circuit1116, small intestine tissue 1106, and liver tissue 1108 correspond to alymphatic system and filter waste and other materials from the firstcircuit 1114.

In the system 1100, each of the cell culture vessels used to implementthe system 1100, provide a top flow and a bottom flow, similar to thecell culture vessel 630 illustrated in FIG. 6D. For example, in the cellculture vessels corresponding to the alveolar tissue 1104 and thetracheobronchial tissue 1102, the cells are provided nutrients throughfluid from the first circuit 1114, which flows through the lower chamberof the cell culture vessels. In the top chamber of the cell culturevessels, the alveolar tissue 1104 and the tracheobronchial tissue 1102are exposed to oxygen. Exposure to oxygen on one side and the fluid ofthe first circuit 1114 on the other, enables the cells of the alveolartissue 1104 and the tracheobronchial tissue 1102 to oxygenate the fluidwhile also removing CO₂.

The bottom flows in the cell culture vessels, which correspond to thesmall intestine tissue 1106 and the liver tissue 1108, also originatefrom the first circuit 1114. As described above, fluid from the firstcircuit 1114 is used to supply the respective tissue with nutrients. Inthe cell culture vessels that correspond to the small intestine tissue1106 and the liver tissue 1108, the top flow is a component of the flowfrom the second circuit 1116. In addition to receiving nutrients fromthe fluid of the first circuit 1114, the small intestine tissue 1106 andthe liver tissue 1108 filter the fluid of the first circuit 1114 andtransfer the filtered waste to the fluid of the second circuit 1116,where it can be removed from the system 1100.

By culturing organ specific tissue types within a biomimetic environment(e.g., within a cell culture vessel as described above wherein thetemperature, humidity, and other parameters mimic in vivo conditions)and interconnecting each of the organ systems in a physiologicallymeaningful way, experiments can be conducted on in vitro cells thatsubstantially mimic the responses of in vivo cell populations. Forexample, a predetermined dose of a drug can be introduced to the system1100 through the drug delivery system 1120. Starting at the drugdelivery system 1120, the first circuit 1114 of the system 1100transports the drug to each of the organ systems 1102, 1104, 1106, and1108. The arrows 1250 illustrate the path taken by drug through thefirst circuit 1114. The cells uptake the drug as it flows through thefirst circuit 1114. Additionally, some of the drug is filtered out ofthe fluid of the first circuit 1114 as it circulates through the system1100. For example, the alveolar tissue 1104 may remove some of the drugas an off gas when the alveolar cells remove CO₂ from the fluid of thefirst circuit 1114. The liver tissue 1108 may also filter the drug outof the fluid of the first circuit 1114 and then transfer the drug to thefluid of the second circuit 1116.

As the drug flows through the system 1100, a number of measurements canbe made. For example, a user may monitor the pH of the fluid in thefirst circuit 1114 to determine if the drug is causing the fluid tobecome basic or acidic. A user may sample the waste collected in thefluid of the second circuit 1116 to determine if the drug dosage is toohigh. For example, a user may perform experiments wherein the drugdosage is lowered to the point where the drug is substantially notpresent in the fluid of the second circuit 1116. In someimplementations, a substantial amount of drug in the fluid of the secondcircuit 1116 indicates that too much drug is being introduced into thesystem 1100.

In some implementations, the user may temporally remove one of the cellculture vessels corresponding to one of the tissue systems and examinethe cells in the cell culture vessel with the above describedmicroscope. For example, the user may examine the cells with amicroscope to determine if the drug is causing damage to the cells. Insome implementations, the user can examine cells within a cell culturevessel without removing the cell culture vessel from the cell cultureplatform.

The invention may be embodied in other specific forms without departingfrom the spirit or essential characteristics thereof. The forgoingimplementations are therefore to be considered in all respectsillustrative, rather than limiting of the invention.

What is claimed:
 1. A modular device for culturing cells, the devicecomprising: a control plate, a first cell culture vessel configured toculture a first type of cells; a second cell culture vessel configureddifferently than the first cell culture vessel and configured to culturea second type of cells; a fluid routing plate configured i) toreversibly receive the first and second cell culture vessels such thatthe first and second cell culture vessels are separately removable fromthe fluid routing plate, and ii) to couple to the control plate, whereinthe fluid routing plate defines a set of channels therethrough to routea fluid flow between the first cell culture vessel and the second cellculture vessel; and a plurality of actuators within the control plate toselectively block a passage of a fluid through a first fluid flowcircuit comprising a first portion of the set of channels between thefirst and second cell culture vessels and selectively enable the passageof the fluid through a second fluid flow circuit comprising a secondportion of the set of channels between the first and second cell culturevessels.
 2. The device of 1, wherein the control plate defines a secondset of channels therethrough to route the fluid flow to at least one ofthe first cell culture vessel and the second cell culture vessel.
 3. Thedevice of claim 1, wherein the first and second cell culture vesselseach further comprise a fluid inlet port and a fluid outlet portconfigured to reversibly and fluidically couple the first and secondcell culture vessels to respective channels in the set of channels. 4.The device of claim 1, wherein the plurality of actuators are configuredto control the state of valves within the fluid routing plate to controlflow rates of the fluid flow through the fluid routing plate.
 5. Thedevice of claim 4, wherein the plurality of actuators are configured toclose the valves by deforming a control membrane within the valves, suchthat the membrane substantially impedes the flow of fluid through achannel in the fluid routing plate.
 6. The device of claim 1, whereinthe plurality of actuators are configured to introduce a predeterminedamount of an agent into the fluid flow between the first and second cellculture vessels.
 7. The device of claim 6, wherein the agent is one of amedication and a toxin.
 8. The device of claim 1, wherein a plurality ofactuators are configured to withdraw fluid samples from one of the firstand second cell culture vessels.
 9. The device of claim 1, wherein thefirst type of cells corresponds to a portion of a first organ system andthe second type of cells corresponds to a portion of a second organsystem.
 10. The device of claim 9, wherein the first organ system isdifferent than the second organ system and each of the first organsystem and the second organ system correspond to one of a kidney, alung, and a liver.
 11. The device of claim 1, wherein each of theplurality of actuators is one of a pneumatic actuator, anelectro-mechanical actuator, and a valve.
 12. The device of claim 1,wherein the control plate further comprises at least one sensor tomeasure a parameter in at least one of the first and second cell culturevessels.
 13. The device of claim 1, wherein the first and second cellculture vessels are disposable.
 14. The device of claim 1, wherein thecontrol plate is configured to withstand a sterilization process. 15.The device of claim 1, wherein first cell culture vessel furthercomprises a first membrane to support the first type of cells and thesecond cell culture vessels comprises a second membrane to support thesecond type of cells.